HEAMATOLOGY ASSIGNMENT

NAME: ADESANMI ADEMOLA O.

STUDENT NUMBER: 221506462

Practical 1
1) a. There should be no stain precipitate present on smear.
b. Stain should not be too dark or too pale.
c. An ideal slide is neither too thin nor too thick.
d. It should end about two-thirds to three-fourths of the way down the slide
2) May-grunewald and giemsa or Wright stain, group- Romanowsky stain
3) Results of practical
a) Relative WCC = Number of cells counted/ 100* 100
Relative WCC = 100/ 100* 100
Relative WCC = 100 %

Absolute WCC = Total WCC x Relative WCC

Absolute WCC = 7.2 x 100%
Absolute WCC = 720 x10^9/L
b) Sickle cells and target cells present

c) Metamyelocytes and Band neutrophils present

d) Giant Platlets present

4) Correct WBC = Total WBC/ (100 + NRBC) *100
= 7.2/ (100+12) *100
= 6.4x10^9/L
5) Possible causes
a) Neutrophilia: Rheumatoid Arthritis and Pregnancy
b) Neutropaenia: HIV and Tuberculosis
c) Lymphocytosis: Hepatitis and Hepatitis
d) Lymphocytopaenia: AIDS and COVID19
e) Monocytosis: Mononucleosis and Rupus
f) Eosinophilia: Parasitic and fungal disease
g) Basophilia: Myelofibrosis and Polycythemia vera
6) A. V. Hoffbrand, Paul A. H. Moss 2016. Hoffbrand’s Essential Haematology, Seventh Edition. Wiley
Blackwell.

Practical 2
1.)
2.) The basic principle of flow cytometry is the single-file passage of cells in front of a laser for
detection, counting, and sorting. The laser excites fluorescently labeled cell components, which
then emit light with a range of wavelengths. The quantity and kind of cells present in the
specimen are then determined by measuring the fluorescence. Finally, we examine the antigens
that are located on the cell. On the surface of every one of our cells are proteins. If our cell
contains that antigen, we add an antibody that has been labeled with a fluorescent marker, and
the antibody will bind to the cell and release light. The presence of the antigen on the cell is
shown by the presence of light.
3.)
CD 34- negative
HLA-DR- negative
CD 13- positive
CD 33- positive
CD 45- positive
CD 117- positive
4.)
a) MYELOPEROXIDASE: Myeloperoxidase in the leukocyte granules oxidizes substrates from an
insoluble blue/brown derivative to a colorless form when hydrogen peroxide is present.
Substrates include benzidine, 3,3′-diaminobenzidine, and p-phenylenediamine
dihydrochloride.
b) NEUTROPHIL ALKALINE PHOSPHATASE: Neutrophilic-Leukocyte The diazotization-coupling
concept is the foundation of alkaline phosphatase stain. Alkaline Phosphatase converts
naphthol AS-BI Phosphoric Acid into -naphthol at pH 9.2–9.8. In the cytoplasm, an insoluble
colored precipitate will be formed by the stable diazonium salt and naphthol.
c) SUDAN BLACK: Sudan Black is a somewhat basic dye that reacts with acidic groups in
complex lipids to stain them as well. a substitute for the Sudan Black B stain.
5.) Fluorescent in situ hybridization: The basic idea is to use a nucleic acid probe to hybridize
nuclear DNA from either interphase cells or metaphase chromosomes attached to a microscopic
slide. The probes are either directly labeled by incorporating a fluorophore or indirectly by
incorporating a hapten. Following denaturation, the labeled probe and the target DNA are
combined, allowing complementary DNA sequences to anneal.
6.)
i) A brown cytoplasmic granulation that is darker than the background staining of red blood
cells is visible in the cell precursors.
 ii) Sudan Black: At the location of granule formation, mature myeloid precursors exhibit
intracellular staining that is black and dark blue.
iii) NAP: Mature neutrophils' secondary granules exhibit a blue granular response in the smear.
7.)
a) Relative WCC = Number of cells counted/ 100* 100
Relative WCC = 3/ 100* 100
Relative WCC = 3%
b) Our haemoglobin level is low, causing anaemia, and our red blood cells are very few and
poikilocytic.
c) Eosinophilic big immature lymphocytes are the type of white cells seen on the ALL slide. The
white cells seen on the APL slide are myeloblasts, which lack granules and have basophilic
cytoplasm and a bilobed nucleus.

d) Because of the extremely low platelet count, the slides demonstrate thrombocytopenia.
e) Because ALL slides have thrombocytopenia due to the low platelet count (75 x 109/L),
Because WCC is high (35x 109/L), ALL slides have leucocytosis.
There are not many red cells on the slide because Hb is low (8g/dL).
8.) Auer rods are used to distinguish between AML and ALL.
9.) A. V. Hoffbrand, Paul A. H. Moss 2016. Hoffbrand’s Essential Haematology, Seventh Edition.
Wiley Blackwell.

Practical 3
1.)
i) Relative WCC = Number of cells counted/ 100* 100
Relative WCC = 50/100*100
Relative WCC = 50%
ii) Rouleaux production in red cells under anisopoikylotic conditions.
 iii) White cells are neutrophils with hypersegmented nuclei, some with band cells, and others
with vacuoles and myelocytes.
iv) Platelets come in a variety of sizes, shapes, and sizes.
v) The patient has CML stage 1 since the slide shows enormous proliferation of granulocytic
cells (neutrophil, Eosinophil, and basophil) and mature neutrophils and myelocytes.
2)
a) PCR: BCR-ABL mRNA
b) Cytogenetics: Philadelphia chromosome
c) FISH: BCR-ABL gene
3)
a) Relative WCC = number of cells counted/ 100*100
Relative WCC = 100/100*100
Relative WCC = 100%
b) The majority of the red cells on the slide are microcytes in a pilocytic state.
c) White cells are mostly lymphocytes and smudge cells, which are shattered lymphocytes during
slide preparation.
d) Platelets are scarce and tiny.
4) Hairy cell leukemia (Hoffbrand, 2016.pp. 202-210):
• Has cytoplasmic projection
• Has a high nucleus-to-cytoplasm ratio
• The nucleus is placed eccentrically
• Chromatin is fine
5) A. V. Hoffbrand, Paul A. H. Moss 2016. Hoffbrand’s Essential Haematology, Seventh Edition. Wiley
Blackwell.

Practical 4
1.) Protein Electrophoresis
Under the influence of an electrical current, serum electrophoresis can separate the various
components of blood proteins into bands or zones based on their electrical charge. Protein
zones are segregated from neighboring zones. The stains then highlight these zones.
2.)

3.)
It has a round eccentric nucleus
It has an abundant basophilic cytoplasm
It has a perinuclear halo
Clockface chromatin is also present
4.)
i) Relative WCC = Number of cells counted/ 100* 100

Relative WCC= 21/100* 100

Relative WCC = 21%

ii) The red cells have a rouleaux formation and a changing size center pallor. Some red cells are
macrocytic (bigger than regular red cells), whilst others are microcytic. Poikilocytosis is
minor among the red cells.
iii) The white cells contain neutrophils with multilobed nuclei, as well as a few monocytes with
eccentric nuclei and lymphocytes with a high nucleus to cytoplasmic ratio.
iv) The morphology of platelets is normal. There are no large or unusual platelets on the slide.
5.)
a) Red cells have minimal rouleaux development and all exhibit minimal anisocytosis.
b) White cell-plasma cells dominate the smear, with an eccentric nucleus and basophilic
cytoplasm. There are also lymphocytes with a high nucleus-to-cytoplasm ratio.
c) Platelet- There are a few big platelets scattered around the slide.
d) Plasma cell leukaemia is defined by abnormally high numbers of plasma cells in the
blood, whereas multiple myeloma is defined by plasma cell accumulation in the bone
marrow
 6.) A. V. Hoffbrand, Paul A. H. Moss 2016. Hoffbrand’s Essential Haematology, Seventh Edition.
Wiley Blackwell.

Practical 5
1)
a) Red cells are poikilocytic because they have a variety of forms. Some are teardrop-shaped,
while others appear undeveloped or juvenile. Other red cells are target cells because they
have a round shape with a circular core pallor. A nucleated red cell is also present.
b) White cells feature a non-segmented nucleus with granules (eosinophils), a banded and
segmented nucleus, and a high nucleus to cytoplasmic ratio (lymphocytes).
c) Platelets- there are big and unusual platelets on the smear.
d) Bone marrow is typically unavailable due to a dry tap.
e) Three probable myelofibrosis genetic mutations
(1) CALR
(2) JAK2V617F
(3) MPL
2)
I) Red cells: the majority of them have typical forms, with only a few having aberrant
morphology.
II) White cells: There are fewer neutrophils with non-segmented nuclei. They contain
fewer segments than mature neutrophils.
III) Platelets - There are numerous normal, tiny, and unusual platelets on the slide.
IV) Increased big hyperlobulated megakaryotes will be found in bone marrow.
3)
a) Have normocytic normochromic red blood cells
b) Have an abundance of normal, big, and unusual platelets
c) Have neutrophil leucocytosis
d) Have mild fibrosis
e) Have hypercellular bone marrow
4) A. V. Hoffbrand, Paul A. H. Moss 2016. Hoffbrand’s Essential Haematology, Seventh Edition. Wiley
Blackwell